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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: FEBS J. 2014 Jul 28;281(17):3816–3827. doi: 10.1111/febs.12905

Fig. 1. Sestrin2 physically associates with ULK1 and p62.

Fig. 1

(A-D) ULK1 and p62 are associated with Sestrin2 in cells. HEK293 cells were transfected with plasmids expressing indicated proteins. HA-tagged proteins were immunopurified (IP) from whole cell lysates (WCL) and subjected to immunoblotting (IB). FIP200 and Atg13 are regulatory subunits of ULK1, and Raptor was used as a negative control. (E,F) ULK1 and p62 bind to Sestrin2 in vitro. HEK293 cells were transfected with plasmids expressing HA-tagged ULK1 (E) or p62 (F). Cell lysates were incubated with GST-Sestrin2 recombinant protein expressed and purified from E. coli, and immunoprecipitated using anti-HA antibody. Pulled-down GST-Sestrin2 proteins were assayed by immunoblotting.